CN102747132A - Aerobic bacterial culture solution for machines - Google Patents
Aerobic bacterial culture solution for machines Download PDFInfo
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- CN102747132A CN102747132A CN2012102540240A CN201210254024A CN102747132A CN 102747132 A CN102747132 A CN 102747132A CN 2012102540240 A CN2012102540240 A CN 2012102540240A CN 201210254024 A CN201210254024 A CN 201210254024A CN 102747132 A CN102747132 A CN 102747132A
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Abstract
The invention discloses an aerobic bacterial culture solution for machines, comprising a nutrient substance needed by bacterial growth, an anticoagulant and an antagonistic antibiotics reagent, wherein the concentration ratio of the three components is 52:3:10; compared with traditional blood bacterium-increasing broth and the culture medium in blood microorganism culture tools such as a manual bidirection aerobic culture flask, the culture solution has high bacterium propagation speed so as to shorten blood detection time; in addition, the bacterium metabolic product can be transformed to a chemical signal capable of being identified by an instrument by reaction with bicarbonate in the culture solution, so as to increase the blood detection accuracy, therefore, the culture solution has wide application prospect in the field of clinical blood examination.
Description
Technical field
The invention belongs to the blood testing technical field, refer more particularly to a kind of machine and use the aerobic-type inoculum.
Background technology
Full-automatic aerobic Blood culture bottle is a kind of novel microorganism culture systems that is applied to clinical medicine check field, is and the matching used microorganism detection reagent of new generation of full-automatic hemoculture appearance.It is made up of three parts such as bottle, solid state sensor and nutrient solutions; Be used in combination with full-automatic hemoculture instrument can rapid detection blood or other samples in whether bacterial infection, fungi and mycobacterium, thereby for the clinical anti-infective therapy that carries out quickly and effectively diagnosis basis is provided.Full-automatic hemoculture system as far back as the eighties of last century the nineties by U.S. company BD; France Mei Liai company etc. begins research and development; Produce in batches at present and sell; China is in the technical know-how state, and external imported product costs an arm and a leg, and brings heavy economical load for the medical institutions and the patient of China.At present China is domestic has only the biological enterprise of minority such as Shanghai Ao Pu biotech company to be engaged in this research, does not form industrialization as yet.Development quality is good, and low-cost automatic culturing bottle is broken international monopoly, not only has good economic benefit, and can reduce the patient burden, reduces medical expense, has social benefit widely.
The nutrient solution majority of one of most important parts is to increase bacterial context soup in the culturing bottle now; Its detection principle provides the necessary nutrient matter of bacterial growth, makes morbific rapid growth of bacteria, thereby metabolic product can be by the physics and the chemical signal of instrument identification through discharging with chemical reagent reaction; Detect the morbific pathogenic agent of blood; Along with developing rapidly of detecting instrument, its precision and sensitivity also improve significantly, and the development of the medicine of antibiotics and generally use; Thereby the variation that nutrient solution also must be adapted; The requirement of nutrient solution now be make vegetative faster, also want in the antagonism with blood in microbiotic, thereby guarantee to detect efficient, save time, the accuracy height.
Summary of the invention
A kind of machine disclosed by the invention is used the aerobic-type inoculum, significantly improves vegetative speed, and in can antagonism with blood in microbiotic, thereby make the speed of bacterial growth fast, shorten the detection time of blood; In addition, opsonigenous substances can through with nutrient solution in supercarbonate reaction be transformed into and can made the blood testing accuracy high by the chemical signal of instrument identification, therefore have application prospects in blood Clinical Laboratory field.
This nutrient solution comprises the required nutritive substance of bacterial growth, anti-coagulant and antagonism antibiotic agent, and the concentration ratio of three kinds of compositions is 52:3:10.The required nutritive substance of bacterial growth can be made up of with VITAMINs and supercarbonate one or more mixtures that peptone, tryptone, Carnis Bovis seu Bubali cream powder, heart and brain soak in powder, the glucose, and its concentration ratio is 26:0.03:0.68; VITAMINs can be made up of one or more mixed vitamins in vitamin A, vitamins D, vitamin E, vitamin B group, the vitamins C; Supercarbonate is made up of in sodium hydrogencarbonate, saleratus, Calcium hydrogen carbonate, the bicarbonate of ammonia one or more; Anti-coagulant can be wherein a kind of arbitrarily of heparin, edetate, Trisodium Citrate BP; Can one or more mix reagents that gather in methyl allylphenol sulfonic acid, para-amino benzoic acid, sal epsom, the absorption microbiotic compound resin with the reagent of microbiotic generation antagonism neutralizing effect in the blood; In addition, when the preparation nutrient solution, also need add salt or acid solution, alkali lye and so in order to the PH environment that is fit to bacterial growth to be provided.
Have the nutritive substance of glucose like above-mentioned formulation selection as bacterial growth; Then in the preparation nutrient solution, should note; Glucose in composition, absorption microbiotic compound resin and the supercarbonate, all weighing mixes heating for dissolving; Need after the dissolving solution is carried out the normal temperature cooling, add glucose and sterile distilled water when treating to cool off slightly; With the nutrient solution autoclaving, be cooled to room temperature simultaneously again; The supercarbonate weighing is dissolved in the sterile distilled water, contain separately in can not leak gas and can autoclaved molten device in, autoclaving is cooled to room temperature; In the aseptic technique platform, both are mixed, regulate pH value to 7.2-7.3, nutrient solution is sub-packed in the Blood culture bottle sterilizes with Autoclave with salt, acid solution or alkali lye; Be cooled to room temperature; Regulate the bottle internal gas pressure to specialized range, seal, nutrient solution can use after batch inspection, the Quality Control.
Embodiment
Prepare the 1500ml Glass Containers of the with closure of two bacterium of going out, the beaker of a 200ml; Accurately weighing peptone 15.00g, tryptone 3.00g, Carnis Bovis seu Bubali cream powder 3.00g, heart and brain soak powder 2.00g, glucose powder 3.00g, Y factor powder 0.03g, sodium bicarbonate powder 0.68g, Trisodium Citrate BP 1.50g, para-amino benzoic acid 0.03g, gather methyl allylphenol sulfonic acid 0.15g, sal epsom 0.20g, sodium-chlor 5.00g, the antibiotic hybrid resin 5g of absorption; Remove glucose, hybrid resin and sodium hydrogencarbonate in the composition; All the other all compositions are packed in the Glass Containers; With sterile distilled water 50ml heating for dissolving, after the dissolving, need solution is carried out the normal temperature cooling fully; Cooling back adds glucose and sterile distilled water slightly, the sterile distilled water to 900 of adding milliliter; With behind the nutrient solution autoclaving 15min, be cooled to room temperature again; Load weighted supercarbonate is dissolved in the sterile distilled water that is contained in the beaker, contain separately in airtight and can autoclaved container in, autoclaving 15min is cooled to room temperature; At the aseptic technique platform both are mixed in the Glass Containers, regulate pH value to 7.2-7.3, add the antibiotic hybrid resin 5g of absorption in addition with hydrochloric acid or sodium hydroxide; Nutrient solution is settled to 1000ml; Nutrient solution is sub-packed in the Blood culture bottle sterilizes, be cooled to room temperature, regulate the bottle internal gas pressure to specialized range with Autoclave; Seal, nutrient solution can use after batch inspection, the Quality Control.
The effect of above-mentioned each composition: peptone and tryptone: mainly as the nitrogenous source of bacterial growth, also can be carbon source, its characteristic is soluble in water, is again two property compounds, has certain shock absorption; The Carnis Bovis seu Bubali cream powder: mainly as the nitrogenous source of bacterial growth, also can be carbon source, it contains the multiple material that can stimulate bacterial growth like creatine, xanthine, uric acid, adenylic acid(AMP), xanthoglobulin, Nucleotide, urea, Stimulina, Beta-alanine, glycogen, phosphohexose, lactic acid, succsinic acid, inorganic salt; Heart and brain soak powder: bacterium provides growth required nutrition.Sodium-chlor: inorganic salt are provided; Glucose: bacterial growth carbon source main provider; Y factor: the growth factor that bacterial growth is provided; Gather methyl allylphenol sulfonic acid: suppress to be unfavorable in the blood material of bacterial growth, like one type in microbiotic, have anticoagulation simultaneously; Para-amino benzoic acid: in the ability with blood in the bacteriostatic action of sulfa drugs; Sal epsom: the bacteriostatic action that can destroy the tsiklomitsin, duomycin, terramycin, polymyxin and the Streptomycin sulphate that exist in the blood; Adsorb antibiotic compound resin: can adsorb multiple microbiotic, particularly β-Nei acyl class microbiotic; Trisodium Citrate BP: prevent that mainly blood coagulation from solidifying; Sodium hydrogencarbonate: can be transformed into meta-bolites in the bacteria growth process and can be utilized acid that glucose produces by red corpuscle survival in the chemical signal of being discerned at the bottom of instrument and the Blood culture bottle, the buffered soln, the cell necessary dioxide gas of growing is provided.
Utilize this nutrient solution to detect the principle of malignant bacteria in the blood: nutrient solution provides the required nutritive substance of malignant bacteria growth in the blood; Need add anti-coagulant and prevent blood coagulation in order to make blood be liquid state all the time; In addition; In order to make bacterial growth rapid, also need add the antagonism antibiotic agent and avoid antibiotic interference in the blood.Bacterium grows in this nutrient solution; It has various meta-bolites; For example: after glucose is utilized by bacterium, can produce pyruvic acid, acetyl methyl carbinol, succsinic acid, lactic acid, (first, second, third, fourth) acid, ethanol, acetate and resolve into carbonic acid gas fully and the proteinic meta-bolites of H2 mainly contains: ammonia, CO2 and various organic acid, amine etc.These meta-bolitess all can make the PH in the nutrient solution change, and following chemical reaction takes place for meta-bolites and sodium hydrogencarbonate:
NaHCO
3,+H
2O
HCO
3 -+Na
+
CO
2+H
2O
H
2CO
3 
?H
++HCO
3 - 
?H
++NaHCO
3+H
2O?
?HCO
3 -+Na
+
Novel color reaction device is all contained in each blood cultivation bottle bottom, in the blood cultivation bottle, has the microorganism growth metabolism to discharge CO
2Or H
+After, the PH in the nutrient solution is changed, the pH that contains Novel color reaction device simultaneously bottom the bottle also changes, and color also changes thereupon, and the change in color signal is sent in the hemoculture appearance, thereby reaches the purpose of detection.
Claims (6)
1. a machine is used the aerobic-type inoculum, it is characterized in that, this nutrient solution comprises the required nutritive substance of bacterial growth, anti-coagulant and antagonism antibiotic agent, and the concentration ratio of three kinds of compositions is 52:3:10.
2. a kind of machine according to claim 1 is used the aerobic-type inoculum; It is characterized in that; The required nutritive substance of said bacterial growth comprises one or more mixtures, VITAMINs and the supercarbonate in peptone, tryptone, Carnis Bovis seu Bubali cream powder, the glucose, and its concentration ratio is 26:0.03:0.68.
3. a kind of machine according to claim 2 is used the aerobic-type inoculum, it is characterized in that, said VITAMINs is made up of one or more mixed vitamins in vitamin A, vitamins D, vitamin E, vitamin B group, the vitamin E.
4. a kind of machine according to claim 2 is used the aerobic-type inoculum, it is characterized in that, said supercarbonate is made up of in sodium hydrogencarbonate, saleratus, Calcium hydrogen carbonate, the bicarbonate of ammonia one or more.
5. a kind of machine according to claim 1 is used the aerobic-type inoculum, it is characterized in that, said anti-coagulant is a kind of in heparin, edetate, the Trisodium Citrate BP.
6. a kind of machine according to claim 1 is used the aerobic-type inoculum, it is characterized in that, said antagonism antibiotic agent is one or more mix reagents that gather in methyl allylphenol sulfonic acid, para-amino benzoic acid, sal epsom, the absorption microbiotic compound resin.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102540240A CN102747132A (en) | 2012-07-23 | 2012-07-23 | Aerobic bacterial culture solution for machines |
| CN2013103075999A CN103343157A (en) | 2012-07-23 | 2013-07-22 | Bacterial culture solution for detecting pathogenic bacteria in blood |
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| CN2012102540240A CN102747132A (en) | 2012-07-23 | 2012-07-23 | Aerobic bacterial culture solution for machines |
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| CN2013103075999A Pending CN103343157A (en) | 2012-07-23 | 2013-07-22 | Bacterial culture solution for detecting pathogenic bacteria in blood |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106222236A (en) * | 2016-07-27 | 2016-12-14 | 郑州点石生物技术有限公司 | Microorganism detection reagent and preparation method thereof in blood |
| CN112831539A (en) * | 2021-01-27 | 2021-05-25 | 浙江夸克生物科技有限公司 | In-vitro detection kit for aerobic microorganisms |
| CN115369147A (en) * | 2022-09-21 | 2022-11-22 | 浙江夸克生物科技有限公司 | Culture solution of aerobic microorganisms and in-vitro detection kit thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112063505A (en) * | 2020-06-15 | 2020-12-11 | 天津市儿童医院 | Bacteria culture bottle containing beta-lactamase and preparation method thereof |
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| US4217411A (en) * | 1978-12-28 | 1980-08-12 | Le Frock Jack L | Novel enrichments for blood culture media |
| CN101089190A (en) * | 2006-06-16 | 2007-12-19 | 阚方琦 | Nutrition blood culture medium |
| CN101144068B (en) * | 2007-09-03 | 2013-06-05 | 王惠萱 | Fast dissolution preparation method for blood culture medium |
| CN101565734A (en) * | 2008-04-21 | 2009-10-28 | 曲奕 | Neutralization antibiotic nutritional blood culture medium |
| FR2942806B1 (en) * | 2009-03-03 | 2011-09-02 | Assist Publ Hopitaux De Paris | METHOD FOR IDENTIFYING GERMS IN A LIQUID ENVIRONMENT |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106222236A (en) * | 2016-07-27 | 2016-12-14 | 郑州点石生物技术有限公司 | Microorganism detection reagent and preparation method thereof in blood |
| CN112831539A (en) * | 2021-01-27 | 2021-05-25 | 浙江夸克生物科技有限公司 | In-vitro detection kit for aerobic microorganisms |
| CN115369147A (en) * | 2022-09-21 | 2022-11-22 | 浙江夸克生物科技有限公司 | Culture solution of aerobic microorganisms and in-vitro detection kit thereof |
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| CN103343157A (en) | 2013-10-09 |
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Application publication date: 20121024 |